Department of Biology, Queen's University, Kingston, Ontario, Canada.
Centre for Biologics Evaluation Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Ontario, Canada.
J Exp Bot. 2017 Dec 16;68(21-22):5857-5869. doi: 10.1093/jxb/erx399.
Phosphoenolpyruvate carboxylase (PEPC) is an important regulatory enzyme situated at a key branch point of central plant metabolism. Plant genomes encode several plant-type PEPC (PTPC) isozymes, along with a distantly related bacterial-type PEPC (BTPC). BTPC is expressed at high levels in developing castor oil seeds where it tightly interacts with co-expressed PTPC polypeptides to form unusual hetero-octameric Class-2 PEPC complexes that are desensitized to allosteric inhibition by L-malate. Analysis of RNA-Seq and microarray transcriptome datasets revealed two distinct patterns of tissue-specific BTPC expression in vascular plants. Species such as Arabidopsis thaliana, strawberry, rice, maize, and poplar mainly exhibited pollen- or floral-specific BTPC expression. By contrast, BTPC transcripts were relatively abundant in developing castor, cotton, and soybean seeds, cassava tubers, as well as immature tomato, cucumber, grape, and avocado fruit. Immunoreactive 118 kDa BTPC polypeptides were detected on immunoblots of cucumber and tomato fruit extracts. Co-immunoprecipitation established that as in castor, BTPCs physically interact with endogenous PTPCs to form Class-2 PEPC complexes in tomato and cucumber fruit. We hypothesize that Class-2 PEPCs simultaneously maintain rapid anaplerotic PEP carboxylation and respiratory CO2 refixation in diverse, biosynthetically active sinks that accumulate high malate levels.
磷酸烯醇式丙酮酸羧化酶(PEPC)是一种位于植物中心代谢分支点的重要调节酶。植物基因组编码几种植物型 PEPC(PTPC)同工酶,以及一种远缘相关的细菌型 PEPC(BTPC)。BTPC 在发育中的蓖麻籽油种子中高水平表达,与共表达的 PTPC 多肽紧密相互作用,形成不寻常的异八聚体类 2 PEPC 复合物,对 L-苹果酸的变构抑制不敏感。RNA-Seq 和微阵列转录组数据集的分析揭示了血管植物中 BTPC 表达的两种不同组织特异性模式。拟南芥、草莓、水稻、玉米和杨树等物种主要表现出花粉或花特异性 BTPC 表达。相比之下,BTPC 转录本在发育中的蓖麻、棉花和大豆种子、木薯块茎以及未成熟的番茄、黄瓜、葡萄和鳄梨果实中相对丰富。在黄瓜和番茄果实提取物的免疫印迹中检测到免疫反应性 118 kDa BTPC 多肽。共免疫沉淀确立了与蓖麻中的情况一样,BTPC 与内源性 PTPC 物理相互作用,在番茄和黄瓜果实中形成类 2 PEPC 复合物。我们假设类 2 PEPC 同时在积累高苹果酸水平的各种生物合成活跃的汇中维持快速的回补性 PEP 羧化作用和呼吸 CO2 重固定作用。